Mannose 6-phosphate receptor and sortilin mediated endocytosis of α-galactosidase A in kidney endothelial cells

Prominent vasculopathy in Fabry disease patients is caused by excessive intracellular accumulation of globotriaosylceramide (GL-3) throughout the vascular endothelial cells causing progressive cerebrovascular, cardiac and renal impairments. The vascular lesions lead to myocardial ischemia, atherogenesis, stroke, aneurysm, thrombosis, and nephropathy. Hence, injury to the endothelial cells in the kidney is a key mechanism in human glomerular disease and endothelial cell repair is an important therapeutic target. We investigated the mechanism of uptake of α-galactosidase A (α-Gal A) in renal endothelial cells, in order to clarify if the recombinant enzyme is targeted to the lysosomes via the universal mannose 6-phosphate receptor (M6PR) and possibly other receptors. Immunohistochemical localization of infused recombinant α-Gal A in a renal biopsy from a classic Fabry disease patient showed that recombinant protein localize in the endothelial cells of the kidney. Affinity purification studies using α-Gal A resins identified M6PR and sortilin as α-Gal A receptors in cultured glomerular endothelial cells. Immunohistochemical analyses of normal human kidney with anti-sortilin and anti-M6PR showed that sortilin and M6PR were expressed in the endothelium of smaller and larger vessels. Uptake studies in cultured glomerular endothelial cells of α-Gal A labeled with fluorescence and (125)I showed by inhibition with RAP and M6P that sortilin and M6PR mediated uptake of α-Gal A. Biacore studies revealed that α-Gal A binds to human M6PR with very high affinity, but M6PR also binds to sortilin in a way that prevents α-Gal A binding to sortilin. Taken together, our data provide evidence that sortilin is a new α-Gal A receptor expressed in renal endothelial cells and that this receptor together with the M6PR is able to internalize circulating α-Gal A during enzyme replacement therapy in patients with Fabry disease.     

Metabolism of praziquantel in kingfish Seriola lalandi

Investigations into the metabolism of drugs used in aquatic animal therapy are useful for understanding the mechanisms of xenobiotic transformation systems and can aid the development of dosing regimens. This study investigated the metabolism of the synthetic anthelmintic praziquantel, which has application in helminthiasis treatment for several fish species including kingfish Seriola lalandi, a commercial aquaculture finfish species. At least 7 mono- or dihydroxylated derivatives of the parent compound were identified in kingfish after administration of a 150 mg kg(-1) oral praziquantel dose, paralleling findings in mammals. The structure of one representative mono-hydroxylated species that was prominent in the skin, muscle, liver, kidney and plasma of kingfish was investigated using fragmentation experiments; this revealed that hydroxylation of the parent molecule occurred in the tetrahydroisoquinoline region of praziquantel, analogous with mammalian metabolites, but different to that of the active mammalian metabolite (trans-4-OH-praziquantel). The implications of these findings with regard to biotransformation systems for this drug in mammals and fish are discussed.     

定量电子束显微分析中生物薄样品的质量损失

对多种生物薄样品和标样进行电子探针X射线能谱显微定量分析,分别以电子束轰击后样品的O Kα峰计数和介于4.2-6.2keV区间的连续X－射线计数变化监测质量损失,结果显示样品O Kα峰计数减少幅度大于连续X－射线计数减少幅度,在相同的分析条件下,各样品质量损失程度不相同（P＜0.05）。培养肝癌细胞冷冻干燥超薄切片、明胶冷冻干燥超薄切片、BSA薄膜、氨基塑料超薄切片、红细胞冷冻干燥超薄切片和卵黄高磷蛋白薄膜样品的质量损失分别为33％、28％、26％、18％、13％和13％,以上结果提示：以O Kα峰计数的减少监测样品的质量损失较敏感,在进行生物薄试样定量EPMA时应对各样品的质量损失进行相应校正。     

126 Porphyra Birdiae Neefus et Mathieson (Bangiales, Rhodophyta): A New Species from the Northwest Atlantic

Recent studies combining biochemical, molecular, and traditional morphological and ecological traits have shown that some currently recognized species of the red algal genus Porphyra are actually "form species" or "complexes" comprising several morphologically similar but genetically distinct taxa. Conflicting reports of chromosome numbers and differences in DNA sequences reported for Porphyra purpurea (Roth) C. Agardh have raised suspicion that more than one taxon has been confused under this name in the Northwest Atlantic. We have identified one of these cryptic taxa and have recently described it as a new species, Porphyra birdiae . Like P. purpurea (Roth) C. Agardh, it has an ovate to broadly elongate, foliose blade with reproductive areas segregated by a distinct line into male and female sectors. While reproductive specimens have historically been confused with P. purpurea , non-reproductive specimens of P. birdiae have been incorrectly identified as P. umbilicalis Kützing. Although P. birdiae is morphologically similar to both of these species, sequences of SSU (nuclear small subunit rRNA gene) and rbc L (plastid ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit gene) indicate that it is not closely related to either one. Based on rbc L sequences, P. birdiae is closely related to P. aestivalis Lindstrom et Fredericq, a proposed new species from Alaska.     

THE DISCOVERY OF HALICYSTIS OVALIS (LYNGBYE ARESCHOUG IN NEW ENGLAND1,2

Halicystis ovalis is recorded for the first time on the northeast coast of North America, from 12–24 m on the exposed open coast of New Hampshire.     

The isolation of minimally degraded hyaluronate from rat skin.

Hyaluronate was isolated quantitatively from fresh rat skin by homogenization in an Edebo [J. Biochem. Microbiol. Technol. Eng. 2, (1960) 453-479] press, extraction with buffered saline, selective elution from DEAE-cellulose, and gel chromatography on Sephadex G-200. Traces of a protein contaminant remained.